Structure-function relationship of licorice (Glycyrrhiza glabra) root extract-xanthan/guar gum mixture in a high sugar content system.

BACKGROUND: Foam-gels are one of the most important multicomponent-model systems in aerated confectionery, and an investigation of their microstructure is desirable. In this research, the structure-function relationship of xanthan gum/guar gum (XG/GG) and licorice (Glycyrrhiza glabra) root extract powder (LEP) was investigated in a high-sugar medium. Foam-gel systems were prepared at 4:10% to 8:20% ratios of LEP to biopolymer. RESULTS: The results show that increasing the LEP content reduced both the melting point and enthalpy, probably due to higher overrun and weaker junctions. Boosting the XG/GG ratio led the enhancement of mechanical properties, whereas increasing the LEP concentration weakened all textural parameters, which could be due to the poor structure of the network in the presence of the foaming agent, increased moisture content and overrun. In the whipped mixture samples containing 10 g kg-1 XG/GG, higher foaming capacity was observed. By increasing the level of biopolymers, smaller and more uniform air cells were formed according to a scanning electron microscopical study. At higher concentration of LEP, smaller bubbles and increased porosity were seen, which could be attributed to the availability of surfactant in the interfacial layer. CONCLUSION: Maximum structural strength was achieved at a 4:20 ratio of LEP to XG/GG. In rheological experiments, pseudoplastic behavior was seen in all samples. Generally, this model system can be simulated for other herbal extracts containing natural surfactants such as saponins. Achieving a more detailed understanding of these structures and their interactions could help in formulating novel food products. © 2021 Society of Chemical Industry.

Separation and determination of alkyl sulfate surfactants in wastewater by capillary electrophoresis coupled with contactless conductivity detection after preconcentration by simultaneous adsorption using alumina beads.

The aim of the present study is to determine four anionic alkyl sulfate (AS) surfactants with different alkyl chains, namely, C8, C10, C12, and C14, in wastewater by CE with capacitively coupled contactless conductivity detection (CE-C4 D). The conditions effective for the separation of the four AS surfactants were systematically optimized and found to be in a Tris-His (50 mM/20 mM) BGE solution at a pH of 8.95, using a separation voltage of +15 kV, hydrodynamic injection by siphoning using a 20 cm injection height and an injection time of 20 s. The LODs for C8, C10, C12, and C14 were 2.58, 2.30, 2.08, and 3.16 mg/L, respectively. The conditions used to achieve the simultaneous adsorption and preconcentration of the AS surfactants using Al2 O3 beads were pH of 3 and 0.1 mM NaCl. The adsorption efficiencies were found to be 45.6, 50.8, 81.7, and 99.9%, while the desorption efficiencies reached 66.1, 70.4, 83.9, and 100.0% for C8, C10, C12, and C14, respectively. The concentrations of the AS surfactants in wastewater samples were quantified by CE-C4 D after preconcentration by simultaneous adsorption using Al2 O3 beads. The results obtained from the proposed method were consistent with those obtained by HPLC-MS/MS, with a deviation of less than 15%. Our results indicate that the CE-C4 D performed after preconcentration by an adsorption technique using Al2 O3 beads is a new, inexpensive, and suitable method for quantifying AS surfactants in wastewater samples.

Purification and identification of a surfactin biosurfactant and engine oil degradation by Bacillus velezensis KLP2016.

Engine oil used in automobiles is a threat to soil and water due to the recalcitrant properties of its hydrocarbons. It pollutes surrounding environment which affects both flora and fauna. Microbes can degrade hydrocarbons containing engine oil and utilize it as a substrate for their growth. Our results demonstrated that cell-free broth of Bacillus velezensis KLP2016 (Gram + ve, endospore forming; Accession number KY214239) recorded an emulsification index (E24%) from 52.3% to 65.7% against different organic solvents, such as benzene, pentane, cyclohexane, xylene, n-hexane, toluene and engine oil. The surface tension of the cell-free broth of B. velezensis grown in Luria-Bertani broth at 35 °C decreased from 55 to 40 mN m-1at critical micelle concentration 17.2 µg/mL. The active biosurfactant molecule of cell-free broth of Bacillus velezensis KLP2016 was purified by Dietheylaminoethyl-cellulose and size exclusion chromatography, followed by HPLC (RT = 1.130), UV-vis spectrophotometry (210 nm) and thin layer chromatography (Rf = 0.90). The molecular weight of purified biosurfactant was found to be ~ 1.0 kDa, based on Electron Spray Ionization-MS. A concentration of 1980 × 10-2 parts per million of CO2 was trapped in a KOH solution after 15 days of incubation in Luria-Bertani broth containing 1% engine oil. Our results suggest that bacterium Bacillus velezensis KLP2016 may promise a new dimension to solving the engine oil pollution problem in near future.

Biosurfactant from endophytic Bacillus pumilus 2A: physicochemical characterization, production and optimization and potential for plant growth promotion.

BACKGROUND: Microbial surfactants called biosurfactants, thanks to their high biodegradability, low toxicity and stability can be used not only in bioremediation and oil processing, but also in the food and cosmetic industries, and even in medicine. However, the high production costs of microbial surfactants and low efficiency limit their large-scale production. This requires optimization of management conditions, including the possibility of using waste as a carbon source, such as food processing by-products. This papers describes the production and characterization of the biosurfactant obtained from the endophytic bacterial strain Bacillus pumilus 2A grown on various by-products of food processing and its potential applications in supporting plant growth. Four different carbon and nitrogen sources, pH, inoculum concentration and temperature were optimized within Taguchi method. RESULTS: Optimization of bioprocess within Taguchi method and experimental analysis revealed that the optimal conditions for biosurfactant production were brewer's spent grain (5% w/v), ammonium nitrate (1% w/v), pH of 6, 5% of inoculum, and temperature at 30 °C, leading to 6.8 g/L of biosurfactant. Based on gas chromatography-mass spectrometry and Fourier transform infrared spectroscopy analysis produced biosurfactant was determined as glycolipid. Obtained biosurfactant has shown high and long term thermostability, surface tension of 47.7 mN/m, oil displacement of 8 cm and the emulsion index of 69.11%. The examined glycolipid, used in a concentration of 0.2% significantly enhanced growth of Phaseolus vulgaris L. (bean), Raphanus L. (radish), Beta vulgaris L. (beetroot). CONCLUSIONS: The endophytic Bacillus pumilus 2A produce glycolipid biosurfactant with high and long tem thermostability, what makes it useful for many purposes including food processing. The use of brewer's spent grain as the sole carbon source makes the production of biosurfactants profitable, and from an environmental point of view, it is an environmentally friendly way to remove food processing by products. Glycolipid produced by endophytic Bacillus pumilus 2A significantly improve growth of Phaseolus vulgaris L. (bean), Raphanus L. (radish), Beta vulgaris L. (beetroot). Obtained results provide new insight to the possible use of glycolipids as plant growth promoting agents.

Seeking faster, alternative methods for glycolipid biosurfactant characterization and purification.

Biosurfactants have been investigated as potential alternatives for synthetic surfactants in several areas, for example, in environmental and pharmaceutical fields. In that regard, extensive research has been carried out with sophorolipids and rhamnolipids that also present various biological properties with therapeutic significance. These biosurfactants are obtained as complex mixtures of slightly different molecules, and thus when studying these microbial glycolipids, the ability to identify and purify the produced compounds is of extreme importance. This study aimed to develop improved methodologies for the identification, separation, and purification of sophorolipids and rhamnolipids. Therefore, an ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was modified to ensure faster characterization of both sophorolipids and rhamnolipids, enabling the identification and fragmentation pattern description of 10 and 13 congeners, respectively. The separation and purification of these biosurfactants was achieved with novel reversed-phase solid-phase extraction methods guaranteeing the isolation of different glycolipids, including those considered for their significant biological activity (e.g. antimicrobial, anticancer). It was possible to isolate sophorolipids and rhamnolipids with purity of 94% and 99%, respectively. The methods presented herein can be easily implemented and are expected to make purification of these biosurfactants easier, facilitating the study of their individual properties in further works.

Marine Microbial-Derived Antibiotics and Biosurfactants as Potential New Agents against Catheter-Associated Urinary Tract Infections.

Catheter-associated urinary tract infections (CAUTIs) are among the leading nosocomial infections in the world and have led to the extensive study of various strategies to prevent infection. However, despite an abundance of anti-infection materials having been studied over the last forty-five years, only a few types have come into clinical use, providing an insignificant reduction in CAUTIs. In recent decades, marine resources have emerged as an unexplored area of opportunity offering huge potential in discovering novel bioactive materials to combat human diseases. Some of these materials, such as antimicrobial compounds and biosurfactants synthesized by marine microorganisms, exhibit potent antimicrobial, antiadhesive and antibiofilm activity against a broad spectrum of uropathogens (including multidrug-resistant pathogens) that could be potentially used in urinary catheters to eradicate CAUTIs. This paper summarizes information on the most relevant materials that have been obtained from marine-derived microorganisms over the last decade and discusses their potential as new agents against CAUTIs, providing a prospective proposal for researchers.

Toxicity Profiling of Biosurfactants Produced by Novel Marine Bacterial Strains.

Surface active agents (SAAs), currently used in modern industry, are synthetic chemicals produced from non-renewable sources, with potential toxic impacts on humans and the environment. Thus, there is an increased interest for the identification and utilization of natural derived SAAs. As such, the marine environment is considered a promising source of biosurfactants with low toxicity, environmental compatibility, and biodegradation compared to their synthetic counterparts. MARISURF is a Horizon 2020 EU-funded project aiming to identify and functionally characterize SAAs, derived from a unique marine bacterial collection, towards commercial exploitation. Specifically, rhamnolipids produced by Marinobacter MCTG107b and Pseudomonas MCTG214(3b1) strains were previously identified and characterized while currently their toxicity profile was assessed by utilizing well-established methodologies. Our results showed a lack of cytotoxicity in in vitro models of human skin and liver as indicated by alamar blue and propidium iodide assays. Additionally, the use of the single gel electrophoresis assay, under oxidative stress conditions, revealed absence of any significant mutagenic/anti-mutagenic potential. Finally, both 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonicacid) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) cell-free assays, revealed no significant anti-oxidant capacity for neither of the tested compounds. Consequently, the absence of significant cytotoxicity and/or mutagenicity justifies their commercial exploitation and potential development into industrial end-user applications as natural and environmentally friendly biosurfactants.

Optimized pulsed electric field-assisted extraction of biosurfactants from Chubak (Acanthophyllum squarrosum) root and application in ice cream.

BACKGROUND: In this study, a face-centered central composite design was applied to optimize pulsed electric field parameters (voltage: 1, 4, 7 kV cm-1 ; pulse number: 10, 65, 120) for the extraction of natural saponins from Chubak root. Data analysis showed that increasing the voltage from 1 to 4 kV cm-1 and pulse number from 10 to 65 increased foaming ability (FA) and emulsion stability, and decreased foam density (FD), foam stability (FS) and lightness, due to the improved extraction of saponins. RESULTS: Whereas, an opposite trend was observed for FA, FD and FS on increasing the voltage from 4 to 7 kV cm-1 as a result of more impurities being extracted. Furthermore, the Chubak root extract (CRE) (0, 1.5, 3.0 and 4.5 g kg-1 ) obtained under the optimized conditions (voltage of 6.4 kV cm-1 and pulse number of 80) was used in ice cream formulation because of its ability to reduce surface tension. Based on the results, the samples containing higher amounts of CRE showed higher viscosity, consistency coefficient, overrun, melting resistance and creaminess, as well as lower values of flow behavior index, hardness, adhesiveness, coarseness and coldness. This could be related to the increased water retention, improved whipping ability, greater fat destabilization and smaller ice crystals. Although more bitterness was perceived as a result of an increase in the level of CRE, it had no negative effect on the overall acceptance assessed by trained sensory panelists. CONCLUSIONS: The results of this study briefly support the conclusion that CRE has a very high potential for use as a foaming, emulsifying and stabilizing agent to improve the quality of ice cream. © 2020 Society of Chemical Industry.

Production of Mannosylerythritol Lipids (MELs) to be Used as Antimicrobial Agents Against S. aureus ATCC 6538.

Antimicrobial resistance (AMR) is a current major health issue, both for the high rates of resistance observed in bacteria that cause common infections and for the complexity of the consequences of AMR. Pathogens like Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumoniae, Mycobacterium tuberculosis among others are clear examples of antibiotic-resistant threats. Biosurfactants have recently emerged as a potential new generation of anti-adhesive and anti-biofilm agents; mannosylerythritol lipids (MELs) are biosurfactants produced by a range of fungi. A range of structural variants of MELs can be formed and the proportion of each isomer in the fermentation depends on the yeast used, the carbon substrate used for growth and the duration of the fermentation. In order to allow assessment of the possible functions of MELs as antimicrobial molecules, small quantities of MEL were produced by controlled fermentation. Fermentations of the yeast Pseudozyma aphidis using rapeseed oil as a carbon source yielded up to 165 gMELs/kgSubstrate. The MELs formed by this strain was a mixture of MEL-A, MEL-B, MEL-C and MEL-D. The MELs produced were tested against S. aureus ATCC 6538 on pre-formed biofilm and on co-incubation biofilm experiments on silicone discs; showing a disruption of biomass, reduction of the biofilm metabolic activity and a bacteriostatic/bactericidal effect confirmed by a release of oxygen uptake [Formula: see text], the reduction of citrate synthase activity and scanning electron microscopy. The results show that MELs are promising antimicrobial molecules for biomedical technological applications that could be studied in detail in large-scale systems and in conjunction with animal tissue models.

Biochemical characterization of a surfactant-stable keratinase purified from Proteus vulgaris EMB-14 grown on low-cost feather meal.

OBJECTIVES: The bioaccumulation of keratinous wastes from poultry and dairy industries poses a danger of instability to the biosphere due to resistance to common proteolysis and as such, microbial- and enzyme-mediated biodegradation are discussed. RESULTS: In submerged fermentation medium, Proteus vulgaris EMB-14 utilized and efficiently degraded feather, fur and scales by secreting exogenous keratinase. The keratinase was purified 14-fold as a monomeric 49 kDa by DEAE-Sephadex A-50 anion exchange and Sephadex G-100 size-exclusion chromatography. It exhibited optimum activity at pH 9.0 and 60 °C and was alkaline thermostable (pH 7.0-11.0), retaining 87% of initial activity after 1 h pre-incubation at 60 °C. The Km and Vmax of the keratinase with keratin azure were respectively 0.283 mg/mL and 0.241 U/mL/min. Activity of P. vulgaris keratinase was stimulated by Ca2+, Mg2+, Zn2+, Na+ and maintained in the presence of some denaturing agents, except ß-mercaptoethanol while Cu2+ and Pb2+ showed competitive and non-competitive inhibition with Ki 6.5 mM and 17.5 mM, respectively. CONCLUSION: This purified P. vulgaris keratinase could be surveyed for the biotechnological transformation of bioorganic keratinous wastes into valuable products such as soluble peptides, cosmetics and biodegradable thermoplastics.

Formulation of Creams Containing Spirulina Platensis Powder with Different Nonionic Surfactants for the Treatment of Acne Vulgaris.

Natural products used in the treatment of acne vulgaris may be promising alternative therapies with fewer side effects and without antibiotic resistance. The objective of this study was to formulate creams containing Spirulina (Arthrospira) platensis to be used in acne therapy. Spirulina platensis belongs to the group of micro algae and contains valuable active ingredients. The aim was to select the appropriate nonionic surfactants for the formulations in order to enhance the diffusion of the active substance and to certify the antioxidant and antibacterial activity of Spirulina platensis-containing creams. Lyophilized Spirulina platensis powder (SPP) was dissolved in Transcutol HP (TC) and different types of nonionic surfactants (Polysorbate 60 (P60), Cremophor A6:A25 (CR) (1:1), Tefose 63 (TFS), or sucrose ester SP 70 (SP70)) were incorporated in creams as emulsifying agents. The drug release was evaluated by the Franz diffusion method and biocompatibility was tested on HaCaT cells. In vitro antioxidant assays were also performed, and superoxide dismutase (SOD) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) assays were executed. Antimicrobial activities of the selected compositions were checked against Staphylococcus aureus (S. aureus) and Cutibacteriumacnes (C. acnes) (formerly Propionibacterium acnes) with the broth microdilution method. Formulations containing SP 70 surfactant with TC showed the most favorable dissolution profiles and were found to be nontoxic. This composition also showed significant increase in free radical scavenger activity compared to the blank sample and the highest SOD enzyme activity was also detected after treatment with the cream samples. In antibacterial studies, significant differences were observed between the treated and control groups after an incubation time of 6 h.

Production and investigation of the physico-chemical properties of MEL-A from glycerol and coconut water.

This work reports the production of MEL-A using coconut water as the carbon source. Proximate analysis of coconut water indicated the presence of nutrients necessary for growth of the organism and production of desired metabolite. The amount of MEL produced using coconut water was 3.85 g/L (± 0.35) with 74% of it being MEL-A when compared to 2.58 g/L (± 0.15) with 60% being MEL-A using glycerol, a conventional carbon source. MEL-A from coconut water consisted of 38.1% long-chain saturated fatty acids (C16:0 and C18:0) whereas with glycerol it was 9.6%. The critical micellar concentration of the biosurfactant from coconut water was 2.32 ± 0.21 µM when compared to 4.41 ± 0.25 µM from glycerol. The stability of O/W emulsion was reduced by 50% and 90% after incubation for 8 h in the case of MEL-A from coconut water and glycerol respectively when compared to synthetic surfactant, Tween-20. MEL-A from both the sources exhibited free radical scavenging activity (DPPH assay) in a dose-dependent manner wherein MEL-A from coconut water showed two fold higher activity than the other. The interaction of coconut water MEL-A with DPPC for drug encapsulation applications was also studied. The DSC measurements showed the differences in the interaction of drugs with DPPC/MEL-A liposome. The differences were also observed in the solubility of drugs after encapsulation with DPPC/MEL-A liposome.

Effects of Monoacyltrehalose Fraction of Rhodococcus Biosurfactant on the Innate and Adaptive Immunity Parameters In Vivo.

The biosurfactant monoacyltrehalose fraction isolated from Rhodococcus ruber IEGM 231 actinobacterium suppresses antibody production, bactericidal potential, and production of IL-1ß by mouse peritoneal cells after intraperitoneal and intramuscular injection and stimulates the production of IL-10 after intraperitoneal injection. The data of in vitro experiments attest to an important role of bacterial glycolipids in the regulation of the functions of splenocytes and peritoneal macrophages.

Bubble-Nucleation-Based Method for the Selective and Sensitive Electrochemical Detection of Surfactants.

We present the first bubble-nucleation-based electrochemical method for the selective and sensitive detection of surfactants. Our method takes advantage of the high surface activity of surfactant analyte to affect the electrochemical bubble nucleation and then transduces the change in nucleation condition to electrochemical signal for determining the surfactant concentration. Using this method, we demonstrate the quantitation of perfluorinated surfactants in water, a group of emerging environmental contaminants, with a remarkable limit of detection (LOD) down to 30 µg/L and a linear dynamic range of over 3 orders of magnitude. With the addition of a preconcentration step, we have achieved the LOD: 70 ng/L, the health advisory for perfluorooctanesulfonate (PFOS) and perfluorooctanoic acid (PFOA) in drinking water established by the U.S. Environmental Protection Agency. The experimental results are in quantitative agreement with our theoretical model derived from classical nucleation theory. Our method also exhibits an exceptional specificity for the surfactant analytes even in the presence of 1000-fold excess of nonsurfactant interference. This method has the potential to be further developed into a universal electrochemical detector for surfactant analysis because of its simplicity and the surface-activity-based detection mechanism.

Inhibition of pathogenic bacterial biofilms on PDMS based implants by L. acidophilus derived biosurfactant.

BACKGROUND: Lactobacillus spp. predominantly shows its presence as a normal mucosal flora of the mouth and intestine. Therefore, the objective of our research is to investigate the in-vitro conditions for the prospective of medically valuable biosurfactants (BSs) derived from Lactobacillus spp. Biosurfactant (BS) obtained from Lactobacillus spp. exhibit antibiofilm and antiadhesive activity against broad range of microbes. In the present study we investigated the production, purification and properties of key components of the cell-associated-biosurfactant (CABS) from Lactobacillus acidophilus NCIM 2903. RESULTS: Extracted, purified, freeze-dried CABS shows reduction in surface tension (SFT) of phosphate buffer saline (PBS @pH 7.0) from 71 to 26 mN/m and had a critical micelle concentration (CMC) of 23.6 mg/mL. The CABS showed reduction in interfacial tension (IFT) against various hydrocarbons and had effective spreading capability as reflected through the decrease in contact angle (CA) on different surfaces (polydimethylsiloxane - PDMS, Teflon tape, glass surface, polystyrene film and OHP sheet). The anionic nature of CABS displayed stability at different pH and temperatures and formed stable emulsions. Thin layer chromatography (TLC) and Fourier transform infrared spectroscopy (FTIR) revealed CABS as glycolipoprotein type. The Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE) showed presence of multiple bands in a molecular range of 14.4 to 60 kDa, with prominent bands of 45 kDa. The CABS has significant antiadhesion and antibiofilm activity against tested bacterial strains. CONCLUSION: The current challenging situation is to develop methods or search for the molecules that will prevent the formations of biofilm on medical bioimplants of PDMS based materials. These findings are supportive for the use of Lactobacilli derived BS as potential antiadhesive agent on various surfaces of biomedical devices.

Antioxidant, antibacterial, and anti-adhesive activities of biosurfactants isolated from Bacillus strains.

Biosurfactants are surface-active compounds that display a range of physiological functions. The present study investigated the antioxidant, antimicrobial, and anti-adhesive or anti-biofilm potential of biosurfactants isolated from Bacillus subtilis VSG4 and Bacillus licheniformis VS16. The antioxidant activity of the biosurfactants was studied in vitro using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and hydroxyl radicals. At 5 mg/mL of the biosurfactant concentration, the scavenging of DPPH and hydroxyl radicals was found to be between 69.1-73.5% and 63.3-69.8%, respectively. The biosurfactants also displayed significant antibacterial activities against both Gram-positive and Gram-negative bacteria. The anti-adhesive activities of the biosurfactants were evaluated against Staphylococcus aureus ATCC 29523, Salmonella typhimurium ATCC 19430, and Bacillus cereus ATCC 11778. The biosurfactants exhibited anti-adhesive activity, even at concentrations of 3-5 mg/mL. Moreover, both biosurfactants displayed notable anti-biofilm activities with a biofilm eradication percentage ranging from 63.9 to 80.03% for VSG4 biosurfactant, and from 61.1-68.4% for VS16 biosurfactant. Furthermore, VSG4 biosurfactant exhibited emulsification and surface tension stability over a wide range of pH (4-10) and temperature up to 100 °C. These results show that VSG4 and VS16 biosurfactants can be potentially used as natural antioxidants, antimicrobials, and/or anti-adhesive agents for food and biomedical applications.

Antimicrobial, anti-adhesive and anti-biofilm potential of biosurfactants isolated from Pediococcus acidilactici and Lactobacillus plantarum against Staphylococcus aureus CMCC26003.

Biosurfactants (BS) are amphipathic compounds produced by diverse groups of microorganisms exhibiting various biological activities. The current study aimed to assess antimicrobial, anti-adhesive and anti-biofilm activities of BS isolated from lactic acid bacteria (LAB), including Pediococcus acidilactici and Lactobacillus plantarum against Staphylococcus aureus CMCC 26003 in vitro. Cell-bound BS from both Pediococcus acidilactici and Lactobacillus plantarum were extracted, and their surface activities were evaluated by oil spreading assay. As quantified by crystal violet method, BS inhibited adhesion and biofilm formation of Staphylococcus aureus in a dose-dependent manner. The above findings were further supported by results of scanning electron microscopy. These two kinds of BS affect expressions of biofilm-related genes (cidA, icaA, dltB, agrA, sortaseA and sarA) and interfere with the release of signaling molecules (AI-2) in quorum sensing systems. Biological activities observed for BS produced by tested LAB suggest prospects for their use against Staphylococcus aureus biofilm-related infections.

Interfacial Activity of Phasin PhaF from Pseudomonas putida KT2440 at Hydrophobic-Hydrophilic Biointerfaces.

Phasins, the major proteins coating polyhydroxyalkanoate (PHA) granules, have been proposed as suitable biosurfactants for multiple applications because of their amphiphilic nature. In this work, we analyzed the interfacial activity of the amphiphilic α-helical phasin PhaF from Pseudomonas putida KT2440 at different hydrophobic-hydrophilic interfacial environments. The binding of PhaF to surfaces containing PHA or phospholipids, postulated as structural components of PHA granules, was confirmed in vitro using supported lipid bilayers and confocal microscopy, with polyhydroxyoctanoate- co-hexanoate P(HO- co-HHx) and Escherichia coli lipid extract as model systems. The surfactant-like capabilities of PhaF were determined by measuring changes in surface pressure in Langmuir devices. PhaF spontaneously adsorbed at the air-water interface, reducing the surface tension from 72 mN/m (water surface tension at 25 °C) to 50 mN/m. The differences in the adsorption of the protein in the presence of different phospholipid films showed a marked preference for phosphatidylglycerol species, such as 1-palmitoyl-2-oleoyl- sn-glycero-3-phosphoglycerol. The PHA-binding domain of PhaF (BioF) conserved a similar surface activity to PhaF, suggesting that it is responsible for the surfactant properties of the whole protein. These new findings not only increase our knowledge about the role of phasins in the PHA machinery but also open new outlooks for the application of these proteins as biosurfactants.

Biosurfactant production by yeasts from different types of soil of the South Shetland Islands (Maritime Antarctica).

AIM: To screen and identify a potential biosurfactant-producing yeast strain isolated from Antarctic soil and to evaluate the fermentation process kinetics of the most promising strain on biosurfactant production using glycerol as carbon source. METHODS AND RESULTS: From the 68 isolated yeast strains, 11 strains were able to produce biosurfactants after Emulsification Index (E.I.) and Drop Collapse tests, reaching an E.I. higher than 10%. Strain 1_4.0 was the best producer, identified as Candida glaebosa based on molecular analysis. Yeast was cultivated in a medium composed of glycerol supplemented with yeast extract for 120 h to determine the process kinetics. The increased C/N ratio affected yeast growth and biosurfactant production. Biosurfactant release was associated with the end of exponential and beginning of the stationary growth phases. Results indicated an E.I. of 30% at the end of the fermentation. CONCLUSIONS: The feasiability of C. glaebosa to produce biosurfactant from a low-cost medium cultivation shows a great impact on the development of bioresource in the Antarctica terrestrial environment. SIGNIFICANCE AND IMPACT OF THE STUDY: Although the diversity of psychrophilic/psychrotolerant micro-organisms from Antarctica has been the preferred subject of study by microbiologists, terrestrial microfungal communities are scarcely investigated and literature about the biotechnological potential of such micro-organisms should cover important biomolecules in addition to cold-adapted enzymes. In the present study, for the first time, the Maritime Antarctica environment was screened as a novel source of biosurfactants produced by micro-organisms.

Characterization of Antifungal Lipopeptide Biosurfactants Produced by Marine Bacterium Bacillus sp. CS30.

This study was initiated to screen for marine bacterial agents to biocontrol Magnaporthe grisea, a serious fungal pathogen of cereal crops. A bacterial strain, isolated from the cold seep in deep sea, exhibited strong growth inhibition against M. grisea, and the strain was identified and designated as Bacillus sp. CS30. The corresponding antifungal agents were purified by acidic precipitation, sequential methanol extraction, Sephadex LH-20 chromatography, and reversed phase high-performance liquid chromatography (RP-HPLC), and two antifungal peaks were obtained at the final purification step. After analysis by mass spectrometry (MS) and tandem MS, two purified antifungal agents were deduced to belong to the surfactin family, and designated as surfactin CS30-1 and surfactin CS30-2. Further investigation showed that although the antifungal activity of surfactin CS30-1 is higher than that of surfactin CS30-2, both of them induced the increased generation of reactive oxygen species (ROS) and caused serious damage to the cell wall and cytoplasm, thus leading to the cell death of M. grisea. Our results also show the differences of the antifungal activity and antifungal mechanism of the different surfactin homologs surfactin CS30-1 and surfactin CS30-2, and highlight them as potential promising agents to biocontrol plant diseases caused by M. grisea.